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Introduction

The gene of an esterase enzyme, called paraoxonase (PON), is a member of a multigene family that comprises three related genes (PON1, PON2 and PON3) with structural homology clustering on the chromosome 7(1)(2)
. Polymorphisms of PON2 gene are associated with risk of cardiovascular diseases such as hypercholesterolemia, noninsulin-dependent diabetes, coronary heart disease (CHD) and myocardial infarction. The codon 192 mutation of PON1 gene is associated with the risk of CHD and has a synergistic effect with the codon 311 (Cys→Ser) polymorphism of PON2 gene. Moreover, the PON2 codon 311 (Cys→Ser) polymorphism might interact with the apoE4 allele to increase the risk of dementia(3)
.

The polymorphism 311 (Cys→Ser) of PON2 gene was determined by allele-specific oligonucleotide PCR assay (ASO-PCR) using primers described by Chen et al.(4)
with patient DNA. This PCR method showed a 96bp DNA fragment for allele C and a 104bp DNA fragment for allele S. In this report, we show the genotyping of allele C in five subjects using both GoTaq® Flexi DNA Polymerase and GoTaq® Hot Start Polymerase (Cat.# M5001).

Materials and Methods

The allele C of polymorphism 311 (Cys→Ser) on PON2 gene was determined by ASO-PCR using the primers in reference 4 with either GoTaq® Flexi DNA Polymerase or GoTaq® Hot Start Polymerase in a 25µl volume. See Table 1 for reaction component volumes and Table 2 for cycling conditions.

Table 1. PCR Mix.

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Table 2. Cycling Conditions.

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PCR products were resolved by electrophoresis on a 10% polyacrylamide (Cat.# V3111) gel and stained with silver nitrate.

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